A new genome engineering method, Prime‑Editing‑Based Inversion with Enhanced Performance (PIE), allows researchers to induce large chromosomal inversions in mammalian cells with improved efficiency and precision. The technique extends prime editing capabilities to structural rearrangements that were previously difficult to model or correct. PIE combines prime editor chemistry with optimized guide RNAs and delivery conditions to invert multi‑megabase regions, enabling functional studies of inversion‑linked diseases and new routes for synthetic chromosome engineering. The method was validated across cell lines and loci, showing higher on‑target performance and reduced off‑target breakage compared with conventional CRISPR‑Cas9 inversion strategies. Researchers anticipate PIE will accelerate functional genomics, disease modeling, and therapeutic approaches that require reordering genomic segments rather than simple base edits.